G protein-coupled receptor signaling : methods and protocols
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Bibliographic Information
G protein-coupled receptor signaling : methods and protocols
(Methods in molecular biology / John M. Walker, series editor, 1947)(Springer protocols)
Humana Press, c2019
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Includes bibliographical references and index
Description and Table of Contents
Description
This detailed volume assembles comprehensive protocols to assist with the study of structural, molecular, cell biological, and in vivo facets of GPCRs, and to enable the development of experimental tools for screening novel GPCR drugs. Sections explore the tweaking of ligands, bioluminescence and FRET approaches, specific GPCR signaling properties, as well as visualization of subcellular compartmentalization. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, G Protein-Coupled Receptor Signaling: Methods and Protocols serves as an ideal reference for life scientists working in a variety of research fields including molecular pharmacology, cell and developmental biology, brain behavior and physiology, drug development and screening.
Chapter 4 is available open access under a CC BY 4.0 license via link.springer.com.
Table of Contents
Part I: Probing Structural Properties of GPCRs
1. Assessment of Conformational State Transitions of Class B GPCRs Using Molecular Dynamics
Chenyi Liao, Victor May, and Jianing Li
2. Molecular Dynamic Simulations to Probe Water Permeation Pathways of GPCRs
Katsufumi Tomobe, Eiji Yamamoto, and Kenji Yasuoka
3. Expression and Purification of a Functional E. coli 13CH3-Methionine-Labeled Thermostable Neurotensin Receptor 1 Variant for Solution NMR Studies
Fabian Bumbak, Ross A.D. Bathgate, Daniel J. Scott, and Paul R. Gooley
4. A Combined Cell-Free Protein Synthesis and Fluorescence-Based Approach to Investigate GPCR Binding Properties
Anne Zemella, Theresa Richter, Lena Thoring, and Stefan Kubick
Part II: Tweaking Ligands for GPCR Studies
5. Furan Cross-Linking Technology for Investigating GPCR-Ligand Interactions
Marleen Van Troys, Willem Vannecke, Christophe Ampe, and Annemieke Madder
6. Optical Regulation of Class C GPCRs by Photoswitchable Orthogonal Remotely-Tethered Ligands
Amanda Acosta-Ruiz, Johannes Broichhagen, and Joshua Levitz
7. Chemoselective Acylation of Hydrazinopeptides to Access Fluorescent Probes for Time-Resolved FRET Assay on GPCRs
Sridevi M. Ramanoudjame, Lucie Esteoulle, Stephanie Riche, Jean-Francois Margathe, Thierry Durroux, Iuliia A. Karpenko, and Dominique Bonnet
Part III: Bioluminescence and Fluorescence Resonance Transfer Energy Approaches
8. Time-Resolved FRET-Based Assays to Characterize G Protein-Coupled Receptor Hetero-Oligomer Pharmacology
Joyce Heuninck, Candide Hounsou, Elodie Dupuis, Eric Trinquet, Bernard Mouillac, Jean-Philippe Pin, Dominique Bonnet, and Thierry Durroux
9. Combining Conformational Profiling of GPCRs with CRISPR/Cas9 Gene Editing Approaches
Kyla Bourque, Dominic Devost, Asuka Inoue, and Terence E. Hebert
10. Measuring GPCR Stoichiometry Using Types -1, -2 and -3 Bioluminescence Resonance Energy Transfer-Based Assays
James H. Felce, John R. James, and Simon J. Davis
11. Combining SRET2 and BiFC to Study GPCR Heteromerization and Protein-Protein Interactions
Amina M. Bagher, Melanie E.M. Kelly, and Eileen M. Denovan-Wright
12. Quantification and Comparison of Signals Generated by Different FRET-Based cAMP Reporters
Andreas Koschinski and Manuela Zaccolo
Part IV: Exploring GPCR Signaling Properties
13. Measuring GPCR-Induced Activation of Protein Tyrosine Phosphatases (PTP) Using In-Gel and Colorimetric PTP Assays
Genevieve Hamel-Cote, Fanny Lapointe, and Jana Stankova
14. Measurement of -Arrestin Recruitment at GPCRs Using the Tango Assay
Genevieve Laroche and Patrick M. Giguere
15. Probing the Interactome of Corticotropin-Releasing Factor Receptor Heteromers Using Mass Spectrometry
Burcu Hasdemir, Juan A. Oses-Prieto, Alma Burlingame, and Aditi Bhargava
Part V: Visualizing Subcellular Compartmentalization of GPCRs
16. Monitoring the Aggregation of GPCRs by Fluorescence Microscopy
Samuel Genier, Jade Degrandmaison, Christine L. Lavoie, Louis Gendron, and Jean-Luc Parent
17. Combining RNAi and Immunofluorescence Approaches to Investigate Post-Endocytic Sorting of GPCRs into Multivesicular Bodies
Xuezhi Li, Stephanie Rosciglione, Andreanne Laniel, and Christine Lavoie
18. Super-Resolution Imaging of G Protein-Coupled Receptors Using Ground State Depletion Microscopy
Fabiana A. Cateano-Crowley, Bryan Heit, and Stephen S.G. Ferguson
19. Analysis of Spatial Assembly of GPCRs Using Photoactivatable Dyes and Localization Microscopy
Kim C. Jonas and Aylin C. Hanyaloglu
Part VI: Investigating In Vivo Functions of GPCRs
20. Optical Modulation of Metabotropic Glutamate Receptor Type 5 In Vivo Using a Photoactive Drug
Marc Lopez-Cano, Joan Font, Amadeu Llebaria, Victor Fernandez-Duenas, and Francisco Ciruela
21. Imaging of Tissue-Specific and Temporal Activation of GPCR Signaling Using DREADD Knock-In Mice
Dmitry Akhmedov, Nicholas S. Kirkby, Jane A. Mitchell, and Rebecca Berdeaux
22. Laser Doppler Flowmetry to Study the Regulation of Cerebral Blood Flow by G Protein-Coupled Receptors in Rodents
Xavier Toussay, Mario Tiberi, and Baptiste Lacoste
23. Preparation of Pulmonary Artery Myocytes and Rings to Study Vasoactive GPCRs
Martha Hinton, Anurag Singh Sikarwar, and Shyamala Dakshinamurti
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